多药耐药相关蛋白（MRP）在人视网膜色素上皮细胞中的表达及其与新型醛糖还原酶抑制剂BAPSG的相互作用。
Expression of multidrug resistance-associated protein (MRP) in human retinal pigment epithelial cells and its interaction with BAPSG, a novel aldose reductase inhibitor.
作者信息
Aukunuru J V, Sunkara G, Bandi N, Thoreson W B, Kompella U B
机构信息
Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha 68198-6025, USA.
出版信息
Pharm Res. 2001 May;18(5):565-72. doi: 10.1023/a:1011060705599.
PURPOSE
The objective of this study was to determine the expression and activity of multidrug resistance-associated protein (MRP) in the retinal pigment epithelial (RPE) cells and to further assess whether BAPSG, a novel anionic aldose reductase inhibitor, interacts with MRP.
METHODS
Functional and biochemical evidence for MRP was obtained in a human retinal pigment epithelial (ARPE-19) cell line and primary cultures of human retinal pigment epithelial (HRPE) cells. Fluorescein accumulation and efflux in the presence and absence of MRP inhibitors was used to obtain functional evidence for MRP. Western blots and RT-PCR were used to obtain biochemical evidence for MRP1. The influence of MRP inhibitors on BAPSG accumulation and efflux in ARPE-19 cells was determined to understand its interaction with MRP.
RESULTS
MRP inhibitors increased fluorescein accumulation and reduced efflux in RPE cells. Both cell types exhibited a 190-kDa western blot band corresponding to MRP1 protein and a 287 bp RT-PCR band corresponding to MRP1 mRNA. MRP inhibitors reduced BAPSG efflux and increased its accumulation in ARPE-19 cells.
CONCLUSIONS
MRP is functionally and biochemically active in human RPE cells. Anionic BAPSG is a likely substrate for MRP.
目的
本研究的目的是确定多药耐药相关蛋白（MRP）在视网膜色素上皮（RPE）细胞中的表达和活性，并进一步评估新型阴离子醛糖还原酶抑制剂BAPSG是否与MRP相互作用。
方法
在人视网膜色素上皮（ARPE - 19）细胞系和人视网膜色素上皮（HRPE）细胞原代培养物中获得MRP的功能和生化证据。在有和没有MRP抑制剂的情况下，通过荧光素积累和流出情况来获得MRP的功能证据。使用蛋白质免疫印迹法（Western blots）和逆转录聚合酶链反应（RT - PCR）来获得MRP1的生化证据。通过测定MRP抑制剂对ARPE - 19细胞中BAPSG积累和流出的影响，以了解其与MRP的相互作用。
结果
MRP抑制剂增加了RPE细胞中荧光素的积累并减少了其流出。两种细胞类型均显示出与MRP1蛋白相对应的190 kDa蛋白质免疫印迹条带和与MRP1 mRNA相对应的287 bp RT - PCR条带。MRP抑制剂减少了ARPE - 19细胞中BAPSG的流出并增加了其积累。
结论
MRP在人RPE细胞中具有功能和生化活性。阴离子BAPSG可能是MRP的底物。
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